A hemostatic composition, its preparation method, and hemostatic products thereof

Abstract

This invention provides a hemostatic composition, its preparation method, and hemostatic products. The raw materials of the hemostatic composition include the following components in parts by weight: 0.01-0.5 parts cannabidiol, 10-150 parts tannic acid-modified carboxymethyl chitosan, and 5-250 parts cross-linked starch. The raw materials of the tannic acid-modified carboxymethyl chitosan include the following components in parts by weight: 45-55 parts carboxymethyl chitosan and 0.5-1.5 parts tannic acid. The raw materials of the cross-linked starch include the following components in parts by weight: 95-105 parts carboxymethyl enzymatically hydrolyzed starch and 2-5 parts sodium trimetaphosphate. This hemostatic composition has good hemostatic effect, and its use in hemostatic products can enable the hemostatic products to have good hemostatic effect.

Description

Technical Field

[0001] This invention relates to the field of hemostatic products technology, and in particular to a hemostatic composition, its preparation method, and hemostatic products. Background Technology

[0002] In many medical scenarios, such as surgery and trauma emergency care, rapid and effective hemostasis is crucial. Traditional hemostatic materials, such as gauze and bandages, while widely used, often fall short in complex bleeding situations, especially in cases of large-area bleeding or where pressure is difficult to apply to the bleeding site. In recent years, with the continuous integration of materials science and biotechnology, novel hemostatic materials have become a research hotspot. However, many existing hemostatic materials still suffer from problems such as insufficient clotting speed, poor biocompatibility, susceptibility to infection, or adverse effects due to residues in the body. Therefore, there is an urgent need to develop a hemostatic product with superior overall performance. Summary of the Invention

[0003] To address the above problems, this invention provides a hemostatic composition. This hemostatic composition has a rapid coagulation effect, and its use in hemostatic products can effectively promote the development of rapid hemostatic products.

[0004] To achieve the above objectives, the present invention adopts the following technical solution:

[0005] A hemostatic composition comprising the following components in parts by weight: 0.01-0.5 parts cannabidiol, 10-150 parts tannic acid-modified carboxymethyl chitosan, and 5-250 parts cross-linked starch; wherein the tannic acid-modified carboxymethyl chitosan comprises the following components in parts by weight: 45-55 parts carboxymethyl chitosan and 0.5-1.5 parts tannic acid; and the cross-linked starch comprises the following components in parts by weight: 95-105 parts carboxymethyl enzymatically hydrolyzed starch and 2-5 parts sodium tripolyphosphate.

[0006] Optionally, the raw material for the carboxymethyl enzymatically hydrolyzed starch includes the following components in parts by weight: 22-30 parts of enzymatically hydrolyzed starch and 15-20 parts of chloroacetic acid.

[0007] Optionally, the raw material for enzymatically hydrolyzing starch includes the following components in parts by weight: 20-25 parts potato starch, 0.2-0.5 parts saccharifying enzyme, and 0.05-0.1 parts α-amylase.

[0008] Optionally, the mass ratio of the saccharifying enzyme to the α-amylase is (2-5):1;

[0009] The sum of the mass fractions of the saccharifying enzyme and the α-amylase is 0.25-0.5 parts.

[0010] Optionally, the raw materials of the hemostatic composition include the following components in parts by weight: 0.05-0.2 parts of cannabidiol, 50-100 parts of tannic acid-modified carboxymethyl chitosan, and 50-150 parts of cross-linked starch;

[0011] The raw material of the tannic acid modified carboxymethyl chitosan includes the following components in parts by weight: 48-52 parts of carboxymethyl chitosan and 0.8-1.2 parts of tannic acid;

[0012] The raw materials for the cross-linked starch include the following components in parts by weight: 98-102 parts of carboxymethyl hydrolyzed starch and 2.5-4 parts of sodium tripolyphosphate;

[0013] The raw material for the carboxymethyl enzymatically hydrolyzed starch comprises the following components in parts by weight: 24-28 parts of the enzymatically hydrolyzed starch and 16-19 parts of the chloroacetic acid;

[0014] The raw materials for the enzymatic hydrolysis of starch include the following components in parts by weight: 21-23 parts of potato starch, 0.3-0.4 parts of saccharifying enzyme, and 0.06-0.09 parts of α-amylase.

[0015] Optionally, the raw materials of the hemostatic composition include the following components in parts by weight: 0.08-0.12 parts of cannabidiol, 70-80 parts of tannic acid-modified carboxymethyl chitosan, and 60-100 parts of cross-linked starch;

[0016] The raw material of the tannic acid modified carboxymethyl chitosan includes the following components in parts by weight: 50 parts of carboxymethyl chitosan and 1 part of tannic acid.

[0017] The raw materials for the cross-linked starch include the following components in parts by weight: 100 parts of carboxymethyl hydrolyzed starch and 3 parts of sodium tripolyphosphate;

[0018] The raw material for the carboxymethyl enzymatically hydrolyzed starch includes the following components in parts by weight: 25 parts of the enzymatically hydrolyzed starch and 18 parts of the chloroacetic acid;

[0019] The raw materials for the enzymatic hydrolysis of starch include the following components in parts by weight: 21 parts of potato starch, 0.32 parts of saccharifying enzyme, and 0.08 parts of α-amylase.

[0020] Optionally, the method is characterized by including the following steps:

[0021] Preparation of the tannic acid-modified carboxymethyl chitosan: The carboxymethyl chitosan is dissolved in water to obtain a carboxymethyl chitosan aqueous solution, wherein the mass percentage of carboxymethyl chitosan in the aqueous solution is 2%-6%; the tannic acid is dissolved in water to obtain a tannic acid aqueous solution, wherein the mass percentage of tannic acid in the aqueous solution is 2%-6%; the tannic acid aqueous solution is slowly added dropwise to the carboxymethyl chitosan aqueous solution, and the reaction is carried out at 55-65℃ for 2-4 hours, maintaining the pH of the reaction solution at 7-9 during the reaction, and air is bubbled in during the reaction. The volume ratio of the tannic acid aqueous solution to the carboxymethyl chitosan aqueous solution is (45-55):1; after the reaction, the reaction solution is dialyzed and then freeze-dried to obtain the tannic acid-modified carboxymethyl chitosan.

[0022] Preparation of the cross-linked starch: The carboxymethyl hydrolyzed starch is dissolved in water to obtain a carboxymethyl hydrolyzed starch solution. The pH of the carboxymethyl hydrolyzed starch solution is adjusted to 9.5-10.5, and then the sodium trimetaphosphate is added to form an aqueous phase. An oil phase matrix and an emulsifier are mixed, wherein the emulsifier accounts for 0.5%-2% of the mass of the oil phase matrix to obtain an oil phase. The aqueous phase is added dropwise to the oil phase, and the reaction is carried out at 55-65℃ for 5-7 hours. After the reaction, the reaction solution is washed with a washing solution including at least one of acetone, petroleum ether, sodium chloride aqueous solution, and ethanol. After washing, the solution is dried at 45-55℃ to obtain the cross-linked starch.

[0023] Preparation of the hemostatic composition: The tannic acid-modified carboxymethyl chitosan and the cross-linked starch are added to a cannabidiol solution and mixed, wherein the cannabidiol solution includes the cannabidiol. Then the solvent in the cannabidiol solution is removed to obtain the hemostatic composition.

[0024] Optionally, the carboxymethyl enzymatically hydrolyzed starch is prepared by the following method:

[0025] Preparation of enzymatically hydrolyzed starch: Potato starch is dissolved in an acetate-sodium acetate buffer solution with a pH of 3.5-4.5, and then saccharifying enzyme and α-amylase are added. The reaction is carried out at 45-55℃ for 6-10 hours. The mass fraction of potato starch is 20-25 parts, the mass fraction of saccharifying enzyme is 0.2-0.5 parts, and the mass fraction of α-amylase is 0.05-0.1 parts. The mass ratio of saccharifying enzyme to α-amylase is (2-5):1, and the sum of the mass fractions of saccharifying enzyme and α-amylase is 0.25-0.5. After the reaction, the product is washed and filtered, and the solid product is vacuum dried at 45-55℃ to obtain enzymatically hydrolyzed starch.

[0026] Sodium hydroxide is dissolved in an ethanol-water solution to obtain a sodium hydroxide solution, wherein the volume percentage of ethanol in the ethanol-water solution is 90%-95%, and the mass percentage of sodium hydroxide in the sodium hydroxide solution is 2%-4%. Enzymatically hydrolyzed starch is added to the sodium hydroxide solution, wherein the mass of the enzymatically hydrolyzed starch and the sodium hydroxide are equal. The reaction is refluxed at 45-55°C for 25-35 minutes. Then, chloroacetic acid is added to the reaction solution, and the reaction is refluxed at 45-55°C for 2-4 hours. The mass fraction of the enzymatically hydrolyzed starch is 22-30 parts, and the mass fraction of the chloroacetic acid is 15-20 parts. After the reaction, the reaction product is washed and filtered, and the solid product is vacuum dried at 45-55°C to obtain carboxymethyl hydrolyzed starch.

[0027] Optionally, the tannic acid aqueous solution is slowly added to the carboxymethyl chitosan aqueous solution at a dropping rate of 1-5 mL / min;

[0028] The oil phase matrix comprises liquid paraffin; the emulsifier comprises Span 80.

[0029] A hemostatic product comprising the hemostatic composition.

[0030] Beneficial effects

[0031] In the hemostatic composition of the present invention, the synergistic effect of the various raw materials results in a hemostatic composition that provides rapid hemostasis and good biocompatibility. Using this composition in hemostatic products can effectively promote the development of rapid hemostatic products. Detailed Implementation

[0032] One embodiment of the present invention provides a hemostatic composition comprising the following components in parts by weight: 0.01-0.5 parts cannabidiol, 10-150 parts tannic acid-modified carboxymethyl chitosan, and 5-250 parts cross-linked starch; the tannic acid-modified carboxymethyl chitosan comprises the following components in parts by weight: 45-55 parts carboxymethyl chitosan and 0.5-1.5 parts tannic acid. The cross-linked starch comprises the following components in parts by weight: 95-105 parts carboxymethyl enzymatically hydrolyzed starch and 2-5 parts sodium tripolyphosphate.

[0033] In the composition of this embodiment, the components work together to achieve a rapid coagulation effect. At the same time, the composition has good antibacterial properties and good biocompatibility.

[0034] Specifically, cannabidiol has certain anti-inflammatory and tissue repair-promoting effects. It can regulate the body's immune response, reduce the damage of inflammatory responses to tissues, and create a favorable environment for wound healing, thereby indirectly helping to stop bleeding and promote wound recovery. At the same time, it has good biocompatibility and does not cause significant immune rejection.

[0035] In tannic acid-modified carboxymethyl chitosan, carboxymethyl chitosan is a water-soluble chitosan derivative with good biocompatibility, biodegradability, and hemostatic properties. It can interact with red blood cells and platelets in the blood, promoting platelet adhesion and aggregation, accelerating the coagulation process, and thus achieving hemostasis. Simultaneously, carboxymethyl chitosan can form a protective film on the wound surface, preventing bacterial infection and promoting wound healing. Tannic acid has an astringent effect, coagulating proteins in wound tissue to form a protective film and reducing blood exudation. Furthermore, tannic acid also has certain antibacterial properties, preventing wound infection. Combined with carboxymethyl chitosan, it further enhances the hemostatic and wound-healing effects. Moreover, the introduction of tannic acid improves the properties of carboxymethyl chitosan, making it more biocompatible.

[0036] In cross-linked starch, carboxymethyl hydrolyzed starch is a product of starch after enzymatic hydrolysis and carboxymethylation modification, exhibiting good water solubility and biocompatibility. It can absorb water from the blood at the wound site, forming a gel-like substance that physically blocks the wound and prevents blood flow. Simultaneously, this gel-like substance provides a moist environment for wound healing, promoting cell migration and tissue repair. Sodium trimetaphosphate, as a cross-linking agent, enables cross-linking reactions between carboxymethyl hydrolyzed starch molecules, forming a three-dimensional network structure. This structure increases the stability and mechanical strength of the starch, allowing it to better perform its hemostatic and protective functions at the wound site, and helps maintain the stability of the local wound microenvironment, thus improving biocompatibility.

[0037] In some embodiments, the mass fraction of cannabidiol in the raw materials of the hemostatic composition may be 0.01 parts, 0.05 parts, 0.1 parts, 0.15 parts, 0.2 parts, 0.25 parts, 0.3 parts, 0.35 parts, 0.4 parts, 0.45 parts, 0.5 parts, etc. It is understood that the mass fraction of cannabidiol can also be appropriately selected within the range of 0.01-0.5 parts.

[0038] In some embodiments, the mass fraction of tannic acid-modified carboxymethyl chitosan in the raw materials of the hemostatic composition can be 10 parts, 20 parts, 30 parts, 40 parts, 50 parts, 60 parts, 70 parts, 80 parts, 90 parts, 100 parts, 110 parts, 120 parts, 130 parts, 140 parts, 150 parts, etc. It is understood that the mass fraction of tannic acid-modified carboxymethyl chitosan can also be appropriately selected within the range of 10-150 parts.

[0039] In some embodiments, the mass fraction of cross-linked starch in the raw materials of the hemostatic composition can be 5 parts, 10 parts, 50 parts, 100 parts, 150 parts, 200 parts, 250 parts, etc. It is understood that the mass fraction of cross-linked starch can also be appropriately selected within the range of 5-250 parts.

[0040] In some embodiments, the raw materials of the hemostatic composition are the following components in parts by weight: 0.01-0.5 parts cannabidiol, 10-150 parts tannic acid-modified carboxymethyl chitosan, and 5-250 parts cross-linked starch. It is understood that the parts by weight of each component can be selected accordingly from the above description.

[0041] In some embodiments, the raw material for carboxymethyl enzymatically hydrolyzed starch includes the following components in parts by weight: 22-30 parts of enzymatically hydrolyzed starch and 15-20 parts of chloroacetic acid.

[0042] Chloroacetic acid is used for carboxymethylation modification of enzymatically hydrolyzed starch. The introduction of carboxymethyl groups gives starch molecules a negative charge. This charge facilitates electrostatic attraction with positively charged components in the blood (such as platelets and clotting factors), thereby accelerating platelet aggregation and clotting factor activation, promoting blood coagulation, and achieving rapid hemostasis. Simultaneously, carboxymethylation improves the water solubility and stability of starch, allowing it to function better at the wound site. The reaction conditions for carboxymethylation modification of enzymatically hydrolyzed starch with chloroacetic acid are relatively mild, and the modified carboxymethylated enzymatically hydrolyzed starch exhibits better hydrophilicity and bioactivity, and is closer to the biomolecular structure of the human body. This structural similarity allows it to interact better with human tissues and cells, reducing foreign body reactions and thus demonstrating good biocompatibility.

[0043] Optionally, the mass fraction of enzymatically hydrolyzed starch in the raw material for carboxymethyl enzymatic hydrolysis can be 22 parts, 23 parts, 24 parts, 25 parts, 26 parts, 27 parts, 28 parts, 29 parts, 30 parts, etc. It is understandable that the mass fraction of enzymatically hydrolyzed starch can also be appropriately selected within the range of 22-30 parts.

[0044] Optionally, the mass fraction of chloroacetic acid in the raw material for carboxymethyl enzymatic hydrolysis of starch can be 15, 16, 17, 18, 19, or 20 parts. Understandably, a suitable selection of chloroacetic acid within the range of 15-20 parts is also possible.

[0045] In some embodiments, the raw materials for enzymatically hydrolyzing starch include the following components in parts by weight: 20-25 parts potato starch, 0.2-0.5 parts saccharifying enzyme, and 0.05-0.1 parts α-amylase.

[0046] Potato starch is a widely available natural polysaccharide. During hemostasis, it acts as a physical barrier, quickly covering the wound surface and initially stopping blood flow. Simultaneously, its granular structure helps adsorb platelets and clotting factors from the blood, promoting platelet aggregation and initiating the clotting process. Furthermore, as a natural biomolecule, potato starch possesses excellent biocompatibility; it is a substance that the human body can metabolize and utilize without causing immune responses or other adverse reactions.

[0047] Two enzymes, saccharifying enzyme and α-amylase, are used to enzymatically hydrolyze potato starch. Saccharifying enzyme acts on the non-reducing ends of starch, gradually hydrolyzing it into sugars such as glucose; α-amylase randomly acts on the α-1,4-glycosidic bonds within the starch, breaking down the long starch chains to generate smaller dextrins and oligosaccharides. Through the synergistic action of these two enzymes, the structure of potato starch is altered, making it easier for the human body to absorb and metabolize. Simultaneously, the enzymatic hydrolysis products may possess better hydrophilicity and bioactivity, enabling them to interact more effectively with blood components and accelerate the blood clotting process. Furthermore, their enzymatic hydrolysis of potato starch is a mild biochemical reaction that does not introduce harmful substances into the human body, and the hydrolysis products are more easily absorbed and utilized, further improving the biocompatibility of the material.

[0048] In some embodiments, the mass fraction of potato starch in the raw material for enzymatically hydrolyzing starch can be 20 parts, 21 parts, 22 parts, 23 parts, 24 parts, 25 parts, etc. It is understood that other suitable choices can be made within the range of 20-25 parts by mass fraction of potato starch.

[0049] In some embodiments, the mass fraction of the saccharifying enzyme in the raw material for enzymatic hydrolysis of starch can be 0.2 parts, 0.3 parts, 0.4 parts, 0.5 parts, etc. It is understood that the mass fraction of the saccharifying enzyme can also be other suitable selections within the range of 0.2-0.5 parts.

[0050] In some embodiments, the mass fraction of α-amylase in the raw material for enzymatic hydrolysis of starch can be 0.05 parts, 0.06 parts, 0.07 parts, 0.08 parts, 0.09 parts, 0.1 parts, etc. It is understood that the mass fraction of α-amylase can also be other suitable selections within the range of 0.05-0.1 parts.

[0051] Furthermore, the mass ratio of saccharifying enzyme to α-amylase is (2-5):1. For example, the mass ratio of saccharifying enzyme to α-amylase can be 2:1, 2.5:1, 3:1, 3.5:1, 4:1, 4.5:1, 5:1, etc. Understandably, other suitable choices can be made within the range of (2-5):1 for the mass ratio of saccharifying enzyme to α-amylase.

[0052] Furthermore, the sum of the mass fractions of the saccharifying enzyme and α-amylase is 0.25-0.5 parts. For example, the sum of the mass fractions of the saccharifying enzyme and α-amylase can be 0.25 parts, 0.3 parts, 0.35 parts, 0.4 parts, 0.45 parts, 0.5 parts, etc. It is understandable that other suitable choices can be made within the range of 0.25-0.5 parts for the sum of the mass fractions of the saccharifying enzyme and α-amylase.

[0053] In some embodiments, the raw materials of the hemostatic composition include the following components in parts by weight: 0.05-0.2 parts cannabidiol, 50-100 parts tannic acid-modified carboxymethyl chitosan, and 50-150 parts cross-linked starch. The raw material of tannic acid-modified carboxymethyl chitosan includes the following components in parts by weight: 48-52 parts carboxymethyl chitosan and 0.8-1.2 parts tannic acid. The raw material of cross-linked starch includes the following components in parts by weight: 98-102 parts carboxymethyl enzymatically hydrolyzed starch and 2.5-4 parts sodium trimetaphosphate. The raw material of carboxymethyl enzymatically hydrolyzed starch includes the following components in parts by weight: 24-28 parts enzymatically hydrolyzed starch and 16-19 parts chloroacetic acid. The raw material of enzymatically hydrolyzed starch includes the following components in parts by weight: 21-23 parts potato starch, 0.3-0.4 parts saccharifying enzyme, and 0.06-0.09 parts α-amylase.

[0054] Further, the raw materials of the hemostatic composition include the following components in parts by weight: 0.08-0.12 parts cannabidiol, 70-80 parts tannic acid-modified carboxymethyl chitosan, and 60-100 parts cross-linked starch. The raw materials of the tannic acid-modified carboxymethyl chitosan include the following components in parts by weight: 50 parts carboxymethyl chitosan and 1 part tannic acid. The raw materials of the cross-linked starch include the following components in parts by weight: 100 parts carboxymethyl enzymatically hydrolyzed starch and 3 parts sodium trimetaphosphate. The raw materials of the carboxymethyl enzymatically hydrolyzed starch include the following components in parts by weight: 25 parts enzymatically hydrolyzed starch and 18 parts chloroacetic acid. The raw materials of the enzymatically hydrolyzed starch include the following components in parts by weight: 21 parts potato starch, 0.32 parts saccharifying enzyme, and 0.08 parts α-amylase.

[0055] Another embodiment of the present invention provides a method for preparing the above-mentioned hemostatic composition, comprising the following steps:

[0056] S101: Preparation of tannic acid-modified carboxymethyl chitosan: Dissolve carboxymethyl chitosan in water to obtain a carboxymethyl chitosan aqueous solution, wherein the mass percentage of carboxymethyl chitosan in the aqueous solution is 2%-6%; dissolve tannic acid in water to obtain a tannic acid aqueous solution, wherein the mass percentage of tannic acid in the aqueous solution is 2%-6%; slowly add the tannic acid aqueous solution dropwise to the carboxymethyl chitosan aqueous solution, and react at 55-65℃ for 2-4 hours, maintaining the pH of the reaction solution at 7-9 during the reaction, and bubbling air during the reaction, with the volume ratio of tannic acid aqueous solution to carboxymethyl chitosan aqueous solution being (45-55):1; after the reaction, dialyze the reaction solution and then freeze-dry it to obtain tannic acid-modified carboxymethyl chitosan. Optionally, the dialysis treatment may include: dialyzing the reaction solution in deionized water for five days using a dialysis bag with a molecular weight cutoff of 1000 Da.

[0057] S102: Preparation of cross-linked starch: Carboxymethyl hydrolyzed starch is dissolved in water to obtain a carboxymethyl hydrolyzed starch solution. The pH of the carboxymethyl hydrolyzed starch solution is adjusted to 9.5-10.5, and then sodium trimetaphosphate is added to form an aqueous phase. An oil phase matrix and an emulsifier are mixed, with the emulsifier accounting for 0.5%-2% of the mass of the oil phase matrix to obtain an oil phase. The aqueous phase is added dropwise to the oil phase, and the reaction is carried out at 55-65℃ for 5-7 hours. After the reaction, the reaction solution is washed with a washing solution, which includes at least one of acetone, petroleum ether, sodium chloride aqueous solution, and ethanol. After washing, the solution is dried at 45-55℃ to obtain cross-linked starch.

[0058] S103: Preparation of hemostatic composition: Tannic acid-modified carboxymethyl chitosan and cross-linked starch are added to a cannabidiol solution and mixed. The cannabidiol solution contains cannabidiol. Then the solvent in the cannabidiol solution is removed to obtain the hemostatic composition.

[0059] Understandably, there are no special restrictions on the order of S101 and S102. S101 can be performed first and then S102, or S102 can be performed first and then S101, or S101 and S102 can be performed simultaneously.

[0060] Alternatively, carboxymethyl enzymatically hydrolyzed starch can be prepared by the following method:

[0061] S201: Preparation of enzymatically hydrolyzed starch: Dissolve potato starch in an acetate-sodium acetate buffer solution with a pH of 3.5-4.5, then add saccharifying enzyme and α-amylase, and react at 45-55℃ for 6-10 hours. The mass fraction of potato starch is 20-25 parts, the mass fraction of saccharifying enzyme is 0.2-0.5 parts, and the mass fraction of α-amylase is 0.05-0.1 parts. The mass ratio of saccharifying enzyme to α-amylase is (2-5):1, and the sum of the mass fractions of saccharifying enzyme and α-amylase is 0.25-0.5. After the reaction, wash the product (e.g., with ultrapure water), filter, and vacuum dry the solid product at 45-55℃ to obtain enzymatically hydrolyzed starch.

[0062] S202: Dissolve sodium hydroxide in an ethanol-water solution to obtain a sodium hydroxide solution. The volume percentage of ethanol in the ethanol-water solution is 90%-95%, and the mass percentage of sodium hydroxide in the sodium hydroxide solution is 2%-4%. Add enzymatically hydrolyzed starch to the sodium hydroxide solution, with equal mass of enzymatically hydrolyzed starch and sodium hydroxide. Reflux at 45-55℃ for 25-35 min. Then add chloroacetic acid to the reaction solution and reflux at 45-55℃ for 2-4 h. The mass fraction of enzymatically hydrolyzed starch is 22-30 parts, and the mass fraction of chloroacetic acid is 15-20 parts. After the reaction, wash the reaction product (e.g., with anhydrous ethanol), filter, and vacuum dry the solid product at 45-55℃ to obtain carboxymethyl hydrolyzed starch.

[0063] Optionally, the tannic acid aqueous solution is slowly added to the carboxymethyl chitosan aqueous solution at a dropping rate of 1-5 mL / min. For example, the dropping rate can be 1 mL / min, 2 mL / min, 3 mL / min, 4 mL / min, 5 mL / min, etc.

[0064] In some embodiments, the oil phase matrix comprises liquid paraffin; the emulsifier comprises Span 80.

[0065] Another embodiment of this application provides a hemostatic product. This hemostatic product comprises the above-described hemostatic composition. This hemostatic product has a good rapid hemostatic effect and good biocompatibility.

[0066] Optionally, hemostatic products include hemostatic dressings.

[0067] Examples 1-5, Comparative Examples 1-5

[0068] The compositions of the hemostatic compositions in the examples and comparative examples are shown in Tables 1-5. Specifically, the mass fractions of the raw material components of the hemostatic compositions are shown in Table 1. The mass fractions of the raw material components of tannic acid-modified carboxymethyl chitosan are shown in Table 2. The mass fractions of the raw material components of cross-linked starch are shown in Table 3. The mass fractions of the raw material components of carboxymethyl enzymatically hydrolyzed starch are shown in Table 4. The mass fractions of the raw material components of enzymatically hydrolyzed starch are shown in Table 5.

[0069] Table 1

[0070]

[0071] Table 2

[0072]

[0073] Table 3

[0074]

[0075] Table 4

[0076]

[0077] Table 5

[0078]

[0079] Test case

[0080] (1) Whole blood coagulation index (BCI) test of hemostatic composition:

[0081] 10 mg of the test composition powder was added to 200 μL of recalcified blood (each 10 μL of blood contained 10 μL of 0.2 mol / L calcium chloride). After incubation at 37 °C with shaking (100 rpm) for 5 min, 10 mL of deionized water was added to dissolve any uncoagulated blood clots. The hemoglobin content of the supernatant was measured at 540 nm using a microplate reader (Tecan, Switzerland). The coagulation index (BCI) was calculated as BCI (%) = (Is - I0) / (Ic - I0) × 100%, where Is represents the absorbance value of the sample, Ic represents the absorbance value of the positive control group (10 mL of deionized water was directly added to 200 μL of recalcified blood), and I0 represents the absorbance value of the blank plate. The test results are shown in Table 6.

[0082] (2) In vitro coagulation time test of hemostatic composition:

[0083] Add 5 mg of the test composition powder sample to a centrifuge tube containing 100 μL of recalcified whole blood. Add 400 μL of PBS at a predetermined time and observe the blood diffusion behavior. The time during which no blood diffuses into the PBS is defined as the clotting time. The test results are shown in Table 6.

[0084] (3) Antibacterial properties test of hemostatic composition:

[0085] Escherichia coli (ATCC8739, Gram-negative bacterium) was selected as the test strain to evaluate the antibacterial properties of the tested composition powder. 100 μL of a powder with a concentration of 10⁷ CFU / mL was used. -1 The *E. coli* solution was added dropwise to the powder gel of the test composition and incubated at 37°C for 2 h. 900 μL of PBS was added and thoroughly mixed. Then, 10 μL of the solution was taken and evenly spread onto a pre-prepared agar plate, and incubated at 37°C for 12 h. Finally, the number of bacterial colonies on the agar plate was observed and recorded. Similarly, 100 μL of *E. coli* solution had a concentration of 10⁷ CFU / mL. -1 The samples were incubated at 37°C for 2 h, then submerged in 900 μL of Luria-Bertani (LB) broth and incubated for another 12 h. Finally, 200 μL of bacterial suspension from each sample tube was transferred to a 96-well plate, and the absorbance at 600 nm was measured using a microplate reader. Each experiment was repeated three times. The sterilization rate was expressed as (ODc-ODs) / ODc×100%, where ODs is the absorbance of the sample and ODc is the absorbance of the blank control without the sample. The test results are shown in Table 6.

[0086] (4) Characterization of the cell compatibility of the hemostatic composition

[0087] After sterilizing each test composition powder by irradiation with ultraviolet light for 24 h, the experiment was conducted. PBS (sample powder to PBS mass ratio of 1:3) was added to the test powder to gel it, and cylinders (diameter 10 mm, height 2 mm) were prepared. After sterilization by soaking in alcohol for 24 h, the cylinders were soaked in culture medium for another 24 h to wash away excess alcohol. L929 cells were seeded at a density of 20,000 cells per well and cultured for 24 h and 48 h, respectively. Cell viability was assessed using the live / dead staining method, and observation and counting were performed using an inverted fluorescence microscope (DMi1, Leica, Germany). The test results are shown in Table 6.

[0088] Table 6

[0089]

[0090] As can be seen from Table 6, when cannabidiol, tannic acid-modified carboxymethyl chitosan, and cross-linked starch are mixed in appropriate mass ratios, the composition can have good hemostatic and antibacterial effects as well as good biocompatibility.

[0091] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

[0092] For those skilled in the art, based on the ideas of this invention, there will be changes in the specific implementation methods and application scope. Therefore, the content of this specification should not be construed as a limitation of this invention.

Claims

1. A method for preparing a hemostatic composition, characterized in that, Includes the following steps: The hemostatic composition comprises the following components in parts by weight: 0.01-0.5 parts cannabidiol, 10-150 parts tannic acid-modified carboxymethyl chitosan, and 5-250 parts cross-linked starch; the tannic acid-modified carboxymethyl chitosan comprises the following components in parts by weight: 45-55 parts carboxymethyl chitosan and 0.5-1.5 parts tannic acid; the cross-linked starch comprises the following components in parts by weight: 95-105 parts carboxymethyl enzymatically hydrolyzed starch and 2-5 parts sodium tripolyphosphate. Preparation of the tannic acid-modified carboxymethyl chitosan: Dissolve the carboxymethyl chitosan in water to obtain a carboxymethyl chitosan aqueous solution, wherein the mass percentage of the carboxymethyl chitosan in the carboxymethyl chitosan aqueous solution is 2%-6%; dissolve the tannic acid in water to obtain a tannic acid aqueous solution, wherein the mass percentage of the tannic acid in the tannic acid aqueous solution is 2%-6%; slowly add the tannic acid aqueous solution dropwise to the carboxymethyl chitosan aqueous solution, react at 55-65℃ for 2-4 hours, maintain the pH of the reaction solution at 7-9 during the reaction, bubble air in during the reaction, and the volume ratio of the tannic acid aqueous solution to the carboxymethyl chitosan aqueous solution is (45-55):1; after the reaction, dialyze the reaction solution and then freeze-dry to obtain the tannic acid-modified carboxymethyl chitosan; Preparation of the cross-linked starch: Dissolve the carboxymethyl enzymatically hydrolyzed starch in water to obtain carboxymethyl... The process involves: 1) preparing a carboxymethyl hydrolyzed starch solution; 2) adjusting the pH of the carboxymethyl hydrolyzed starch solution to 9.5-10.5, then adding the sodium trimetaphosphate and mixing to form an aqueous phase; 3) mixing an oil phase matrix and an emulsifier, wherein the emulsifier accounts for 0.5%-2% of the mass of the oil phase matrix to obtain an oil phase; 4) adding the aqueous phase dropwise into the oil phase and reacting at 55-65℃ for 5-7 hours; 5) washing the reaction solution with a washing solution including at least one of acetone, petroleum ether, sodium chloride aqueous solution, and ethanol, and drying at 45-55℃ to obtain the cross-linked starch; 6) preparing the hemostatic composition by mixing the tannic acid-modified carboxymethyl chitosan and the cross-linked starch in a cannabidiol solution, wherein the cannabidiol solution includes cannabidiol, and then removing the solvent from the cannabidiol solution to obtain the hemostatic composition.

2. The method for preparing the hemostatic composition according to claim 1, characterized in that, The carboxymethyl enzymatically hydrolyzed starch was prepared by the following method: Potato starch was dissolved in an acetate-sodium acetate buffer solution with a pH of 3.5-4.5, and then saccharifying enzyme and α-amylase were added. The reaction was carried out at 45-55℃ for 6-10 hours. The mass fraction of potato starch was 20-25 parts, the mass fraction of saccharifying enzyme was 0.2-0.5 parts, and the mass fraction of α-amylase was 0.05-0.1 parts. The mass ratio of saccharifying enzyme to α-amylase was (2-5):1, and the sum of the mass fractions of saccharifying enzyme and α-amylase was 0.25-0.

5. After the reaction, the product was washed and filtered, and the solid product was vacuum dried at 45-55℃ to obtain the enzymatically hydrolyzed starch. Hydroxide... Sodium is dissolved in an ethanol-water solution to obtain a sodium hydroxide solution, wherein the volume percentage of ethanol in the ethanol-water solution is 90%-95%, and the mass percentage of sodium hydroxide in the sodium hydroxide solution is 2%-4%. Enzymatically hydrolyzed starch is added to the sodium hydroxide solution, wherein the mass of the enzymatically hydrolyzed starch and the sodium hydroxide are equal. The reaction is refluxed at 45-55°C for 25-35 minutes. Then, chloroacetic acid is added to the reaction solution, and the reaction is refluxed at 45-55°C for 2-4 hours. The mass fraction of the enzymatically hydrolyzed starch is 22-30 parts, and the mass fraction of the chloroacetic acid is 15-20 parts. After the reaction, the reaction product is washed and filtered, and the solid product is vacuum dried at 45-55°C to obtain carboxymethyl hydrolyzed starch.

3. The method for preparing the hemostatic composition according to any one of claims 1-2, characterized in that, The tannic acid aqueous solution is slowly added dropwise to the carboxymethyl chitosan aqueous solution at a rate of 1-5 mL / min; the oil phase matrix includes liquid paraffin; and the emulsifier includes Span 80.

4. The method for preparing the hemostatic composition as described in claim 1, characterized in that, The raw materials for the carboxymethyl enzymatically hydrolyzed starch include the following components in parts by weight: 22-30 parts of enzymatically hydrolyzed starch and 15-20 parts of chloroacetic acid.

5. The method for preparing the hemostatic composition according to claim 4, characterized in that, The raw materials for the enzymatic hydrolysis of starch include the following components in parts by weight: 20-25 parts potato starch, 0.2-0.5 parts saccharifying enzyme, and 0.05-0.1 parts α-amylase.

6. The method for preparing the hemostatic composition according to claim 5, characterized in that, The mass ratio of the saccharifying enzyme to the α-amylase is (2-5):1; the total mass of the saccharifying enzyme and the α-amylase is 0.25-0.5 parts.

7. The method for preparing the hemostatic composition as described in claim 2, characterized in that, The raw materials of the hemostatic composition include the following components in parts by weight: 0.05-0.2 parts of cannabidiol, 50-100 parts of tannic acid-modified carboxymethyl chitosan, and 50-150 parts of cross-linked starch; the raw materials of the tannic acid-modified carboxymethyl chitosan include the following components in parts by weight: 48-52 parts of carboxymethyl chitosan and 0.8-1.2 parts of tannic acid; the raw materials of the cross-linked starch include the following components in parts by weight: 98-102 parts of carboxymethyl enzymatically hydrolyzed starch and 2.5-4 parts of sodium trimetaphosphate; the raw materials of the carboxymethyl enzymatically hydrolyzed starch include the following components in parts by weight: 24-28 parts of enzymatically hydrolyzed starch and 16-19 parts of chloroacetic acid; the raw materials of the enzymatically hydrolyzed starch include the following components in parts by weight: 21-23 parts of potato starch, 0.3-0.4 parts of saccharifying enzyme, and 0.06-0.09 parts of α-amylase.

8. The method for preparing the hemostatic composition as described in claim 2, characterized in that, The raw materials of the hemostatic composition include the following components in parts by weight: 0.08-0.12 parts of cannabidiol, 70-80 parts of tannic acid-modified carboxymethyl chitosan, and 60-100 parts of cross-linked starch; the raw materials of the tannic acid-modified carboxymethyl chitosan include the following components in parts by weight: 50 parts of carboxymethyl chitosan and 1 part of tannic acid; the raw materials of the cross-linked starch include the following components in parts by weight: 100 parts of carboxymethyl enzymatically hydrolyzed starch and 3 parts of sodium trimetaphosphate; the raw materials of the carboxymethyl enzymatically hydrolyzed starch include the following components in parts by weight: 25 parts of enzymatically hydrolyzed starch and 18 parts of chloroacetic acid; the raw materials of the enzymatically hydrolyzed starch include the following components in parts by weight: 21 parts of potato starch, 0.32 parts of saccharifying enzyme, and 0.08 parts of α-amylase.

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